A major project that involves the TIS is the definition of molecular biomarkers for autoimmune and autoinflammatory diseases. In collaboration with Dan Kastner and Ivona Aksentijevich, we have measured serum cytokines levels in patients with a novel autoinflammatory disease caused by mutations in the LUBAC complex ultimately leading to NFkappaB-mediated proinflammatory cytokines overproduction. Furthermore, in collaboration with the Kastner group and Elaine Remmers we have been studying a new Behcets disease-associated loci that we identified using the Immunochip to densely interrogate immune/inflammatory disease loci. We found a noncoding SNP in IL1A associated with disease at genome-wide significance. We found that the disease-associated SNP is also a reported eQTL; the disease risk allele is significantly associated with decreased expression of IL1A in lymphoblastoid cell lines and skin, and the lead disease-associated SNP is also the lead IL1A eQTL SNP. The TIS has been responsible for confirming decreased lipopolysaccharide-stimulated IL1A mRNA expression in healthy control monocytes from disease risk allele homozygotes compared with individuals with 0 or 1 risk allele. Association of the lead disease SNP with IL1B expression was not statistically significant in the published eQTL study. IL-1 is highly expressed in the epidermis and plays an important role in skin barrier functions against pathogens. The manuscript reporting these findings was published in Nature Genetics. With the Goldbach-Mansky group, we studied patients with Chronic Atypical Neutrophilic Dermatosis with Lipodystrophy and Elevated temperature (CANDLE), a Proteasome-Associated Autoinflammatory Syndrome (PRAAS), who were treated with the JAK inhibitor baricitinib. The TIS was responsible for measuring the levels of phosphor STATs in the leukocytes from these patients and correlate these to the pharmacokinetics of the drug. A manuscript is currently under review at the Journal of Clinical Investigation. In collaboration with the Colbert group we have investigated the cytokine profile and the immunologic phenotype of a patient with a de-novo gain-of-function mutation in MyD88, which results in early-onset severe arthritis. A manuscript describing these results is currently being submitted. We continue to collaborate with NHGRI investigators assessing the cytokine-secreting capabilities of patients with Erdheim-Chester Disease (ECD), a rare, non-familial multisystem disorder characterized by proliferation and infiltration of non-Langerhans histocytes into multiple organs. We compared the cytokines secreted by ECD patients to normal controls after stimulation of PBMCs. The TIS has also been investigating novel approaches for the treatment of autoimmune diseases. In collaboration with the O'Shea group, the Kaplan group, and Pfizer (via a CRADA), we are currently evaluating the effects of tofacitinib and second-generation JAK-selective inhibitors on T cells and innate lymphoid cells. We generated induced pluripotent stem cells (iPSC) from a patient with HLA B-27+ Ankylosing Spondylitis and demonstrated their pluripotency. We then used CRISPR/Cas9 gene editing technology to target HLA-B*27:04 through non-homologous end joining by editing the coding sequence at exon 3. RNA-Seq was used to identify in-del mutations, frameshifts, missense mutations, or premature stop codons. Two types of mutations were identified by RNA-Seq on the targeting site of exon 3 of the B27 allele that were predicted to disrupt B27 protein expression. Western Blot results were consistent with the genomic data indicating that B27 protein expression is decreased significantly in the knock-out cell lines. The newly generated isogenic HLA-B27 knock-out IPS cell lines are also in the process of being characterized.